Objective To investigate the effects of Zhuang medicinal thread moxibustion on apoptosis of interstitial cells of Cajal(ICC)in gastric smooth muscle of diabetic gastroparesis(DGP)rats via regulation of the p38MAPK signaling pathway.Methods Totally 60 rats were randomly divided into a blank group(10 rats)and a modeling group(50 rats).DGP models were established in the modeling group via intraperitoneal injection of streptozotocin.50 successfully modeled rats were further randomized into model group,Western medication group,prevention+treatment group,prevention group and treatment group,with 10 rats in each group.The Western medication group was administered mosapride citrate by gavage for 3 weeks starting at week 10;the prevention+treatment group underwent Zhuang medicinal thread moxibustion("Zhongwan",bilateral"Neiguan"and"Zusanli")intervention for 6 weeks starting at week 7;the prevention group received Zhuang medicinal thread moxibustion for 3 weeks starting at week 7;and the treatment group received Zhuang medicinal thread moxibustion for 3 weeks starting at week 10,3 cones per point,once a day;the blank group and model group received handling only.The random blood glucose and body mass of rats were detected,and the gastric emptying rate and intestinal propulsion rate were calculated.HE staining was used to observe the morphology of gastric smooth muscle tissue,TUNEL staining was used to detect the apoptosis of gastric smooth muscle tissue,ELISA was used to detect the contents of Caspase-3,TNF-α,IL-1β and IL-6 in gastric smooth muscle tissue,immunohistochemistry was used to detect the positive expressions of Cx43 and c-kit in gastric smooth muscle tissue,Western blot was used to detect the expressions of p38MAPK,p-p38MAPK,BAX and Bcl-2 protein in gastric smooth muscle tissue.Results Compared with the blank group,the random blood glucose significantly increased in the model group(P<0.01),and the body mass,gastric emptying rate and small intestinal propulsion rate significantly decreased(P<0.01),the blood vessels of gastric tissue were not clear,the surface was not smooth,the mucosal folds were less,the arrangement of gastric smooth muscle cells was disordered,the nuclear membrane was not clear,and there were vacuole like changes between cells,the apoptosis rate significantly increased(P<0.01),the contents of Caspase-3,TNF-α,IL-1β,IL-6 and the protein expressions of p38MAPK,p-p38MAPK and BAX in gastric smooth muscle tissue significantly increased(P<0.01),and the positive expressions of Cx43,c-kit,the protein expression of Bcl-2 and Bcl-2/BAX ratio significantly decreased(P<0.01).Compared with the model group,the random blood glucose of rats in each intervention group decreased,and the body mass,gastric emptying rate and intestinal propulsion rate increased(P<0.01),the blood vessels of gastric tissue were clear,the inner surface of stomach was smooth,the mucosal folds were not rich,no ulcer was found,the shape of gastric smooth muscle cells was complete,the nuclear membrane was clear,and the arrangement of muscle cells was regular,the apoptosis rate significantly decreased(P<0.01),the contents of Caspase-3,TNF-α,IL-1β,IL-6 and the expressions of p38MAPK,p-p38MAPK,BAX protein in gastric smooth muscle tissue decreased(P<0.05,P<0.01),the positive expressions of Cx43,c-kit,the protein expression of Bcl-2 and Bcl-2/BAX ratio increased(P<0.05,P<0.01).The overall effect of the prevention+treatment group was better than that of the Western medication group,prevention group and treatment group(P<0.05,P<0.01).Conclusion Zhuang medicinal thread moxibustion may reduce gastric inflammation and ICC apoptosis in DGP rats by inhibiting p38MAPK signaling pathway,so as to improve DGP gastric motility disorder,and the effect of early intervention is more significant.

Objective:To identify the change trajectory of intrinsic capacity in people aged ≥50 years in Shanghai and explore the impact of intrinsic capacity trajectory change on overall function and dalily life activities in this population.Methods:The longitudinal data from round 1 to 3 Study of Global Ageing and Adult Health in Shanghai were used. The total intrinsic ability scores from five dimensions of cognition, psychology, sensory, vitality and locomotion were calculated. The censored normal model of group-based trajectory was used to identify the trajectory of intrinsic capacity change over time. Linear regression model and multivariate logistic regression model were used to analyse the effects of different levels intrinsic capacity trajectory on the scores of the WHO Disability Assessment Schedule (WHODAS), the activity of daily living (ADL) and the instrumental activities of daily living (IADL).Results:A total of 2 302 study participants aged ≥50 years with 3 round complete data were included in this study, and 3 levels of intrinsic capacity trajectory were identified, low-level trajectory (9.3%), medium-level trajectory (41.7%), and high-level trajectory (49.0%). Compared with the high-level group, the medium-level and low-level groups had higher WHODAS scores, which increased by 3.578 (95% CI: 2.028-5.129) and 12.620 (95% CI: 9.951-15.289), respectively, and those with more severe disability and those in the low-level group were at higher risk for severe difficulty in ADLs ( OR=12.450, 95% CI: 4.310-35.966) and IADLs ( OR=5.479, 95% CI: 1.311-22.904). Conclusions:Heterogeneity in trajectory of intrinsic capacity exists in people aged ≥50 years in Shanghai. Middle-aged and elderly people with low initial level and rapid decline trajectory of intrinsic capacity are at greater risk for the decline of daily life ability and the increase of disability. It is necessary to strengthen the long-term dynamic monitoring and evaluation of the change trajectory of intrinsic capacity in this population.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

Objective:To identify the change trajectory of intrinsic capacity in people aged ≥50 years in Shanghai and explore the impact of intrinsic capacity trajectory change on overall function and dalily life activities in this population.Methods:The longitudinal data from round 1 to 3 Study of Global Ageing and Adult Health in Shanghai were used. The total intrinsic ability scores from five dimensions of cognition, psychology, sensory, vitality and locomotion were calculated. The censored normal model of group-based trajectory was used to identify the trajectory of intrinsic capacity change over time. Linear regression model and multivariate logistic regression model were used to analyse the effects of different levels intrinsic capacity trajectory on the scores of the WHO Disability Assessment Schedule (WHODAS), the activity of daily living (ADL) and the instrumental activities of daily living (IADL).Results:A total of 2 302 study participants aged ≥50 years with 3 round complete data were included in this study, and 3 levels of intrinsic capacity trajectory were identified, low-level trajectory (9.3%), medium-level trajectory (41.7%), and high-level trajectory (49.0%). Compared with the high-level group, the medium-level and low-level groups had higher WHODAS scores, which increased by 3.578 (95% CI: 2.028-5.129) and 12.620 (95% CI: 9.951-15.289), respectively, and those with more severe disability and those in the low-level group were at higher risk for severe difficulty in ADLs ( OR=12.450, 95% CI: 4.310-35.966) and IADLs ( OR=5.479, 95% CI: 1.311-22.904). Conclusions:Heterogeneity in trajectory of intrinsic capacity exists in people aged ≥50 years in Shanghai. Middle-aged and elderly people with low initial level and rapid decline trajectory of intrinsic capacity are at greater risk for the decline of daily life ability and the increase of disability. It is necessary to strengthen the long-term dynamic monitoring and evaluation of the change trajectory of intrinsic capacity in this population.

This study was aimed at investigating the effectiveness of various host nucleic acid removal and non-specific amplifica-tion techniques in animal tissue samples,to increase the accuracy of pathogen identification in tissue samples.Simulated samples were prepared with a mixture of mouse lung tissue homogenates and Klebsiella pneumoniae fluids,and processed with six host nucleic acid removal kits and three non-specific amplification techniques.The effectiveness of each method in removing host DNA and enriching nucleic acids of pathogenic microorganisms was evaluated through real-time fluorescence quantitative PCR and high-throughput se-quencing.For host nucleic acid removal techniques,the method of selective cleavage and quantitative degradation of host DNA(Com-plete5 kit)effectively decreased the host nucleic acid content in tissue samples and increased the relative abundance of pathogen nucleic acids.In contrast,the magnetic bead method for host DNA removal(Next microbiome DNA enrichment Kit kit)was less effec-tive.At lower pathogen concentrations(77 CFU/mL),the Vazyme kit was more effective than the other kits in removing host nucleic acids.Non-specific amplification techniques(MALBAC whole genome amplification,MDA isothermal amplification,and random primer amplification)were not applicable to tissue samples and were not effective in increasing the relative abundance of pathogen nucleic acids.Selective lysis and quantitative degradation of host DNA were suitable for processing tissue samples with high host back-ground and low pathogenic microorganism levels,whereas non-specific amplification methods were not applicable to tissue samples for pre-processing of macro-genome high-throughput sequencing.

Objective To investigate the effects of Zhuang medicinal thread moxibustion on apoptosis of interstitial cells of Cajal(ICC)in gastric smooth muscle of diabetic gastroparesis(DGP)rats via regulation of the p38MAPK signaling pathway.Methods Totally 60 rats were randomly divided into a blank group(10 rats)and a modeling group(50 rats).DGP models were established in the modeling group via intraperitoneal injection of streptozotocin.50 successfully modeled rats were further randomized into model group,Western medication group,prevention+treatment group,prevention group and treatment group,with 10 rats in each group.The Western medication group was administered mosapride citrate by gavage for 3 weeks starting at week 10;the prevention+treatment group underwent Zhuang medicinal thread moxibustion("Zhongwan",bilateral"Neiguan"and"Zusanli")intervention for 6 weeks starting at week 7;the prevention group received Zhuang medicinal thread moxibustion for 3 weeks starting at week 7;and the treatment group received Zhuang medicinal thread moxibustion for 3 weeks starting at week 10,3 cones per point,once a day;the blank group and model group received handling only.The random blood glucose and body mass of rats were detected,and the gastric emptying rate and intestinal propulsion rate were calculated.HE staining was used to observe the morphology of gastric smooth muscle tissue,TUNEL staining was used to detect the apoptosis of gastric smooth muscle tissue,ELISA was used to detect the contents of Caspase-3,TNF-α,IL-1β and IL-6 in gastric smooth muscle tissue,immunohistochemistry was used to detect the positive expressions of Cx43 and c-kit in gastric smooth muscle tissue,Western blot was used to detect the expressions of p38MAPK,p-p38MAPK,BAX and Bcl-2 protein in gastric smooth muscle tissue.Results Compared with the blank group,the random blood glucose significantly increased in the model group(P<0.01),and the body mass,gastric emptying rate and small intestinal propulsion rate significantly decreased(P<0.01),the blood vessels of gastric tissue were not clear,the surface was not smooth,the mucosal folds were less,the arrangement of gastric smooth muscle cells was disordered,the nuclear membrane was not clear,and there were vacuole like changes between cells,the apoptosis rate significantly increased(P<0.01),the contents of Caspase-3,TNF-α,IL-1β,IL-6 and the protein expressions of p38MAPK,p-p38MAPK and BAX in gastric smooth muscle tissue significantly increased(P<0.01),and the positive expressions of Cx43,c-kit,the protein expression of Bcl-2 and Bcl-2/BAX ratio significantly decreased(P<0.01).Compared with the model group,the random blood glucose of rats in each intervention group decreased,and the body mass,gastric emptying rate and intestinal propulsion rate increased(P<0.01),the blood vessels of gastric tissue were clear,the inner surface of stomach was smooth,the mucosal folds were not rich,no ulcer was found,the shape of gastric smooth muscle cells was complete,the nuclear membrane was clear,and the arrangement of muscle cells was regular,the apoptosis rate significantly decreased(P<0.01),the contents of Caspase-3,TNF-α,IL-1β,IL-6 and the expressions of p38MAPK,p-p38MAPK,BAX protein in gastric smooth muscle tissue decreased(P<0.05,P<0.01),the positive expressions of Cx43,c-kit,the protein expression of Bcl-2 and Bcl-2/BAX ratio increased(P<0.05,P<0.01).The overall effect of the prevention+treatment group was better than that of the Western medication group,prevention group and treatment group(P<0.05,P<0.01).Conclusion Zhuang medicinal thread moxibustion may reduce gastric inflammation and ICC apoptosis in DGP rats by inhibiting p38MAPK signaling pathway,so as to improve DGP gastric motility disorder,and the effect of early intervention is more significant.

Objective:To observe the effects of Zhulian stimulant type Ⅰ acupuncture on the expression of brain-derived neurotrophic factor (BDNF) and its receptor TrkB and tissue homogenate cyclic adenosine phosphate (cAMP) in rats with diabetic bladder (DCP); To explore the mechanism of Zhulian stimulant type Ⅰacupuncture on DCP.Methods:Totally 50 SD rats were divided into control group, model group, Western medicine group, ordinary acupuncture group, Zhulian stimulant type Ⅰ acupuncture treatment group (acupuncture treatment group) according to random number table method, with 10 rats in each group. DCP rat model was prepared by intraperitoneal injection of streptozotocin (STZ), except for the control group. The Western medicine group was given mecobalamine for gavage; acupoints of "Zhongji", "Sanyinjiao", "Liechou" and "Taichong" were selected. The ordinary acupuncture group was treated with ordinary acupuncture technique, and the acupuncture treatment group was treated with Zhulian stimulant type Ⅰ acupuncture, 1 time/d, 30 minutes/time. Samples were taken after 4 weeks of treatment. The maximum bladder volume, residual urine volume and wet weight of the bladder were detected. The morphology of rat bladder was observed by HE staining. The expression level of BDNF was detected by immunohistochemistry. The expression of cAMP was detected by Western blot. The level of TrkB was determined by ELISA. Real-time fluorescence quantitative PCR (RT-PCR) was used to detect the mRNA expressions of BDNF and cAMP.Results:Compared with model group, maximum bladder volume, residual urine volume and wet weight of bladder in Western medicine group, ordinary acupuncture group and acupuncture treatment group decreased ( P<0.01), and those in Western medicine group and acupuncture treatment group were lower than those in ordinary acupuncture group ( P<0.01). The expressions of BDNF mRNA and protein, cAMP mRNA and protein in Western medicine group, ordinary acupuncture group and acupuncture treatment group increased ( P<0.05), and the level of TrkB increased, and the Western medicine group and acupuncture treatment group were higher than that in ordinary acupuncture group ( P<0.05). Conclusions:Zhuliping stimulant type Ⅰ acupuncture has a protective effect on the bladder function of diabetic rats. The mechanism may be related to the up-regulation of BDNF and mRNA, TrkB, cAMP and mRNA expressions.

Objective:To investigate dietary patterns of individuals aged ≥50 in Shanghai and analyze their association with frailty.Methods:Using data from the third wave of the Study on Global Ageing and Adult Health in Shanghai conducted between 2018 and 2019. We collected the frequency and average intake of food by the food frequency questionnaire. Factor analysis was used to extract dietary patterns, and a frailty index was constructed using the ratio of the cumulative total score of health deficits to 35 health-related variables considered. We used an ordinal multinomial logistic regression model to analyze the association between dietary patterns and frailty.Results:A total of 3 274 participants aged (67.9±9.2) years were included in the study, including 1 971 (60.2%) men and 1 303 (39.8%) women. We extracted four dietary patterns: high-protein-nuts pattern, potato-bean-vegetable-fruit pattern, poultry-meat pattern, and high-oil-salt pattern. After adjusting for confounding factors, the logistic regression analysis showed that compared with the high-oil-salt pattern, the high-protein-nuts pattern was negatively associated with the risk of higher frailty ( OR=0.743, 95% CI: 0.580-0.951). We did not find an association between dietary patterns and frailty between the different gender groups. In the age group 50-64, the high-protein-nuts and potato-bean-vegetable-fruit patterns were negatively correlated with a higher degree of frailty than the high-oil-salt pattern. In the low-level physical activity group, the high-protein-nuts pattern was negatively correlated with a higher degree of frailty than the high-oil-salt pattern ( OR=0.509, 95% CI: 0.361-0.720). However, we found no significant effect of the high-protein nuts pattern, potato-bean-vegetable-fruit pattern, and poultry-meat pattern on the risk of higher frailty compared to the high-oil-salt pattern in the moderate to high level of physical activity group. Conclusions:Compared to the high-oil-salt pattern, dietary patterns with a higher intake of high-protein nuts, potatoes, legumes, and fruits and vegetables might be associated with a lower risk of higher frailty in residents aged 50-64 years of age than with a high oil and salt pattern. At the same time, it may have a more significant protective effect in people with lower physical activity levels. It is suggested that a diet rich in high-protein foods, nuts, potatoes, beans, vegetables, and fruits may help reduce and delay the risk of frailty.

Objective:To understand the association between vitamin D level and grip strength in people aged ≥50 years in Shanghai.Methods:Data were obtained from the WHO's Study on Global Ageing and Adult Health in Shanghai during 2018-2019. Logistic regression model was used to analyze the association between vitamin D level and grip strength, and a stratified analysis was conducted for different gender, age and dairy product intake groups. Restricted cubic spline was used to evaluate the dose-response association between vitamin D level and low grip strength.Results:A total of 4 391 participants were included in the study, including 2 054 men (46.8%), with an average age of (67.02±8.81) years. And 1 421 individuals (32.4%) had low grip strength; 1 533 individuals (34.9%) had vitamin D deficiency, and 401 individuals (9.1%) had vitamin D deficiency. After adjusted for confounding factors, the logistic regression results analysis showed that individuals with vitamin D deficiency had a higher risk for low grip strength ( OR=1.41, 95% CI: 1.09-1.83). In men, after adjusting for confounding factors, vitamin D deficiency was positively associated with the risk for low grip strength ( OR=1.67, 95% CI: 1.12-2.50), but there was no significant association between vitamin D level and grip strength in women ( OR=1.30, 95% CI: 0.97-1.74). In age group 60-69 years and ≥80 years, there was significant association between vitamin D deficiency and low grip strength after adjusting for confounding factors ( OR=1.57, 95% CI: 1.05-2.35; OR=2.40, 95% CI: 1.08-5.31). In people who had daily intake of dairy product <250 ml, there was positive association between vitamin D deficiency and low grip strength, but there was no significant association in people who had daily dairy product ≥250 ml after adjusting for confounding factors. The restrictive cubic spline demonstrated that risk of low grip strength might decreased with the increase of vitamin D levels, however, the difference was not significant ( P>0.05). Conclusions:This study demonstrated that there is association between vitamin D level and grip strength. People with vitamin D deficiency have higher risk for low grip strength.